Background: Agro-wastes were used for the production of fibrinolytic enzyme in solid-state fermentation. The
process parameters were optimized to enhance the production of fibrinolytic enzyme from
Bacillus halodurans
IND18 by statistical approach. The fibrinolytic enzyme was purified, and the properties were studied.
Results: A two-level full factorial design was used to screen the significant factors. The factors such as moisture,
pH, and peptone were significantly affected enzyme production and these three factors were selected for
further optimization using central composite design. The optimum medium for fibrinolytic enzyme production
was wheat bran medium containing 1% peptone and 80% moisture with pH 8.32. Under these optimized
conditions, the production of fibrinolytic enzyme was found to be 6851 U/g. The fibrinolytic enzyme was
purified by 3.6-fold with 1275 U/mg specific activity. The molecular mass of fibrinolytic enzyme was
determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis, and it was observed as 29 kDa.
The fibrinolytic enzyme depicted an optimal pH of 9.0 and was stable at a range of pH from 8.0 to 10.0. The
optimal temperature was 60°C and was stable up to 50°C. This enzyme activated plasminogen and also
degraded the fibrin net of blood clot, which suggested its potential as an effective thrombolytic agent.
Conclusions: Wheat bran was found to be an effective substrate for the production of fibrinolytic enzyme. The
purified fibrinolytic enzyme degraded fibrin clot. The fibrinolytic enzyme could be useful to make as an
effective thrombolytic agent.
